3-heterocyclic thiomethyl cephalosporins

ABSTRACT

Cephalosporin compounds having a 3-position substituent of the formula (I) are described: 
     
         --CH.sub.2 --S--Q--(Y).sub.n --P 
    
     wherein Q is a 5- or 6-membered heterocyclic ring, P is a benzene ring substituted by groups R 1  and R 2  which are ortho with respect to one another, wherein R 1  is hydroxy or an in vivo hydrolysable ester thereof and R 2  is hydroxy, an in vivo hydrolysable ester thereof, carboxy, sulpho, hydroxymethyl, methanesulphonamido or ureidos; or P is a group of the formula (II) or (III): ##STR1## wherein M is oxygen or a group NR 3  wherein R 3  is hydrogen or C 1-4  alkyl; said ring P being further optionally substituted; and --(Y) n  -- is a bond or various linking groups or --(Y)-- n  may be such so that rings Q and P are fused. 
     The use of such compounds as antibacterial agents is described as are processes for their preparation and intermediates therefor.

This is a division of application Ser. No. 07/740,420, filed Aug. 5,1991, now U.S. Pat. No. 5,262,410, which is a division of appln. Ser.No. 07/133,482, filed Dec. 15, 1987, now U.S. Pat. No. 5,057,511.

The present invention relates to cephalosporins and in particular tosuch compounds comprising a catechol or related group. This inventionfurther relates to processes for their preparation, to intermediates intheir preparation, to their use as therapeutic agents and topharmaceutical compositions containing them. The compounds of thisinvention are antibiotics and can be used in the treatment of anydisease that is conventionally treated with antibiotics for example inthe treatment of bacterial infection in mammals including humans. Thecompounds of this invention also have non-therapeutic uses as they canbe used in conventional manner in industry for example they can be usedas disinfectants and food preservatives. The compounds of thisinvention, however, are primarily of therapeutic interest as they show adesirable profile of activity and duration in their antibacterialeffect.

Investigation into new cephalosporin derivatives has been intense overthe past 25 years with many thousands of patents and scientific papershaving been published. A particular problem associated with thecommercially available cephalosporins is the lack of potency againststrains of Pseudomonas. The present invention provides cephalosporinderivatives having novel 3-position substituents, which derivativespossess good antibacterial activity and in particular against strains ofPseudomonas.

U.S. Pat. No. 4,278,793 discloses cephalosporins having a 3-positionsubstituent of the formula: --CH₂ Y wherein Y can be the residue of anucleophilic compound, preferably a sulphur, nitrogen or oxygennucleophilic compound. GB1496757 discloses cephalosporins having a3-position substituent of the formula: --CH₂ Y wherein Y can be theresidue of a nucleophilic compound; this specification includes adiscussion of nitrogen, carbon, oxygen and sulphur nucleophiles. Manypossible values of Y are mentioned. These specifications are typical ofmany specifications that describe cephalosporins having nucleophilicmoities linked via a methylene group to the 3-position of acephalosporin. However, although there has been intense research over along period of time, there has been no teaching or suggestion of thecompounds of the present invention. These contain specific ring systemsthat are characterised by having hydroxy groups or related substituentsortho to one another. These hitherto undisclosed ring systems give riseto particularly good activity against strains of Pseudomonas.

Accordingly the present invention provides a cephalosporin compoundhaving a 3-position substituent of the formula (I):

    --CH.sub.2 --S--Q--(Y).sub.n --P                           (I)

wherein Q represents a 5- or 6-membered heterocyclic ring containing 1-4heteroatoms selected from oxygen, nitrogen and sulphur (optionally fusedto a benzene ring or to a further such heterocyclic ring) wherein Qoptionally, where possible, may bear a positive charge and mayoptionally be substituted on an available carbon or nitrogen atom bycarboxy, sulpho, C₁₋₄ alkoxycarbonyl or C₁₋₄ alkyl (which alkyl groupmay itself optionally be substituted by carboxy, sulpho or C₁₋₄alkoxycarbonyl);

P represents:

(i) a benzene ring (optionally fused to a further benzene ring (soforming a naphthyl group) or to a 5 or 6 membered heterocyclic aromaticgroup containing 1, 2 or 3 heteroatoms selected from nitrogen, oxygenand sulphur) said benzene ring (or in the case of naphthyl eitherbenzene ring) substituted by groups R¹ and R² which are ortho withrespect to one another wherein R¹ is hydroxy or an in vivo hydrolysableester thereof and R² is hydroxy, an in vivo hydrolysable ester thereof,carboxy, sulpho, hydroxymethyl, methanesulphonamido or ureido;

(ii) a group of the formula (II): ##STR2## or

(iii) a group of the formula (III): ##STR3## wherein M is oxygen or agroup NR³ wherein R³ is hydrogen or C₁₋₄ alkyl:

ring P (or, in the case wherein ring P is a benzene ring and is fused toanother benzene ring, either benzene ring) is optionally furthersubstituted by C₁₋₄ alkyl, halo, hydroxy, hydroxy C₁₋₄ alkyl, cyano,trifluoromethyl, nitro, amino, C₁₋₄ alkylamino, di-C₁₋₄ alkylamino,amino C₁₋₄ alkyl, C₁₋₄ alkylamino C₁₋₄ alkyl, di-C₁₋₄ alkylamino C₁₋₄alkyl, C₁₋₄ alkanoyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio, C₁₋₄ alkanoyloxy,carbamoyl, C₁₋₄ alkylcarbamoyl, di-C₁₋₄ alkyl carbamoyl, carboxy,carboxy C₁₋₄ alkyl, sulpho, sulpho C₁₋₄ alkyl, C₁₋₄ alkanesulphonamido,C₁₋₄ alkoxycarbonyl, C₁₋₄ alkanoylamino, nitroso, thioureido, amidino,ammonium, mono- , di- or tri-C₁₋₄ alkylammonium pyridinium, or a5-membered heterocyclic ring containing 1 to 4 heteroatoms selected fromoxygen, nitrogen and sulphur which is optionally substituted by 1, 2 or3 C₁₋₄ alkyl or C₁₋ 4 alkoxy groups,

n=0 or 1 such that when n=1 Y represents a covalent bond between Q and Por a C₁₋₄ alkylene group optionally substituted by carboxy or sulpho orY represents a group --(CH₂)_(m) --Y'-- wherein m=1 or 2 and Y' is--O.CO-- or --NH.CO--; and when n=0 Q and P both represent monocyclicrings which are fused on an available carbon--carbon or carbon--nitrogenbond.

In one particular aspect --(Y)_(n) -- is C₁₋₄ alkylene optionallysubstituted by carboxy. In another particular aspect --(Y)_(n) -- is acovalent bond directly linking groups Q and P. In yet another aspect--(Y)_(n) -- is such that rings Q and P are fused to form a bicyclicring system. In a preferred aspect Y is a covalent bond or a methyleneor ethylene linkage.

In one aspect ring P is a benzene ring substituted by groups R¹ and R²as hereinbefore defined. R¹ is hydroxy or an in vivo hydrolysable esterthereof. In vivo hydrolysable esters are those pharmaceuticallyacceptable esters that hydrolyse in the human or animal body to producethe parent hydroxy compound. Such esters can be identified byadministering, e.g. intravenously to a test animal, the compound undertest and subsequently examining the test animal's body fluids. Suitablein vivo hydrolysable esters include C₁₋₆ alkanoyloxy for exampleacetoxy, propionyloxy, pivaloyloxy, C₁₋₄ alkoxycarbonyloxy for exampleethoxycarbonyloxy, phenylacetoxy and phthalidyl.

Conveniently both R¹ and R² have the same value and are both hydroxy orare both in vivo hydrolysable esters, for example they are both acetoxyor both pivaloyloxy.

In a preferred aspect P is a benzene ring optionally fused to anotherbenzene ring so forming a naphthyl group. As stated hereinbefore eitherbenzene group may be substituted by R¹ and R² and by other optionalsubstituents. Particular optional substituents are C₁₋₄ alkyl forexample methyl, ethyl or isopropyl, halo for example chloro, bromo orfluoro, hydroxy, hydroxy C₁₋₄ alkyl for example hydroxymethyl, amino,nitro, C₁₋₄ alkoxy for example methoxy or ethoxy, carboxy C₁₋₄ alkyl forexample carboxymethyl, C₁₋₄ alkanoylamino for example acetamido,trifluoromethyl, carboxy, carbamoyl, cyano, sulpho, C₁₋₄alkanesulphonamido for example methanesulphonamido, C₁₋₄ alkanoyl forexample acetyl, C₁₋₄ alkanoyloxy for example acetoxy or propionoxy andC₁₋₄ alkoxycarbonyl for example methoxycarbonyl. Of these, favouredsubstituents are sulpho, carboxymethyl, methyl, ethyl, methoxy, bromo,chloro, fluoro and nitro.

The skilled man will realise that when P is a benzene ring up to 3optional substituents are possible; when a naphthyl ring is formed moresubstituents are possible and up to 2 or 3 substituents are possiblewith the rings of formulae (II) and (III). In general, we prefer up to 2optional substituents, which may be the same or different.

Particular meanings for ring Q when it is a 5-membered ring arepyrazole, imidazole, thiazole, isothiazole, thiadiazole, triazole andtetrazole. In one aspect Q is imidazole, pyrazole, isothiazole, triazoleor tetrazole. In another aspect Q is thiadiazole or thiazole wherein Yis A C₁₋₄ alkylene group optionally substituted by carboxy or sulpho orY represents a group --(CH₂)_(m) --Y'-- as defined hereinbefore.

In a further aspect ring Q is a 6-membered ring for example pyridine,pyridazine, pyrimidine and pyrazine, in which n=1 and Y may be linked toa carbon atom of Q or to a nitrogen atom, or n=0 and the rings Q and Pare fused. When Y is linked to a carbon atom of Q, or when Q and P arefused, Q may be uncharged, or may bear a positive charge for examplewhen a nitrogen atom of Q is substituted, for example by a C₁₋₄ alkylgroup eg a methyl group. When Y is linked to a nitrogen atom of such agroup Q then Q will bear a positive charge.

Particular meanings for --S--Q-- are rings having the structures set outin formulae VI-XII: ##STR4##

As stated hereinabove the present invention relates to cephalosporinshaving a novel 3-position substituent. A particular class ofcephalosporins within the present invention is that of the formula(XIII): ##STR5## and salts and esters thereof wherein Q, Y, P and n areas hereinbefore defined;

X is sulphur, oxygen, methylene or sulphinyl;

R⁴ is hydrogen, methoxy or formamido; and R⁵ and R⁶ are groups known forsuch positions in the cephalosporin art.

Preferably X is sulphur.

Preferably R⁴ is hydrogen.

R⁵ is for example 2-aminothiazol-4-yl or 2-aminooxazol-4-yl eachoptionally substituted in the 5-position by fluorine, chlorine orbromine, or R⁵ is 5-aminoisothiazol-3-yl, 5-amino-1,2,4-thiadiazol-3-yl,3-aminopyrazol-5-yl, 3-aminopyrazol-4-yl, 2-aminopyrimidin-5-yl,2-aminopyrid-6-yl, 4-aminopyrimidin-2-yl, 2-amino-1,3,4-thiadiazol-5-ylor 5-amino-l-methyl-1,2,4-triazol-3-yl;

R₆ is for example of the formula ═N.O.R⁷ (having the syn configurationabout the double bond) wherein R⁷ is hydrogen, (1-6C)alkyl,(3-8C)cycloalkyl, (1-3C)alkyl(3-6C)cycloalkyl,(3-6C)cycloalkyl(1-3C)alkyl, (3-6C)alkenyl, optionally substituted bycarboxy, (5-8C)cycloalkenyl, (3-6C)alkynyl, (2-5C)alkylcarbamoyl,phenylcarbamoyl, benzylcarbamoyl, (1-4C)alkylcarbamoyl(1-4C)alkyl,di(1-4C)alkylcarbamoyl(1-4C)alkyl, (1-4C)haloalkylcarbamoyl(1-4C)alkyl,(1-3C)haloalkyl, (2-6C)hydroxyalkyl, (1-4C)alkoxy(2-4C)alkyl,(1-4C)alkylthio(2-4C)alkyl, (1-4C)alkanesulphinyl(1-4C)alkyl,(1-4C)alkanesulphonyl(1-4C)alkyl, (2-6C)aminoalkyl, (1-4C)alkylamino(1-6C)alkyl, (2-8C)dialkylamino(2-6C)alkyl,(1-5C)cyanoalkyl, 3-amino-3-carboxypropyl, 2-(amidinothio)ethyl,2-(N-aminoamidinothio)ethyl, tetrahydropyran-2-yl, thietan-3-yl,2-oxopyrrolidinyl, or 2-oxotetrahydrofuranyl, or R⁷ is of the formulaXIV: ##STR6## wherein q is one or two and R⁸ and R⁹ are independentlyhydrogen or C₁₋₄ alkyl; or R⁷ is of the formula XV:

    --CR.sup.10 R.sup.11 --(CH.sub.2).sub.r --COR.sup.12       XV

wherein r is 0-3, R¹⁰ is hydrogen, (1-3C)alkyl or methylthio, R¹¹ ishydrogen (1-3C)alkyl, (3-7C)cycloalkyl, cyano, carboxy,(2-5C)carboxyalkyl or methanesulphonylamino, or R¹⁰ and R¹¹ are joinedto form, together with the carbon to which they are attached, a(3-7C)carbocyclic ring, and R¹² is hydroxy, amino, (1-4C)alkoxy, (1-4C)alkylamino or of the formula NHOR¹³ in which R¹³ is hydrogen or(1-4C)alkyl;

or R⁶ may be of the formula ═CH.R¹⁴ wherein R¹⁴ is hydrogen, halogen,(1-6C)alkyl, (3-7C)cycloalkyl, (2-6C)alkenyl, (3-7C)cycloalkenyl, phenylor benzyl.

Particular meanings for R⁷ are hydrogen, methyl, ethyl, isopropyl,t-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,methylcyclopropyl, methylcyclobutyl, methylcyclopentyl,methylcyclohexyl, cyclopropylmethyl, cyclobutylmethyl,cyclopentylmethyl, allyl, cyclopentenyl, cyclohexenyl, propargyl,methylcarbamoyl, ethylcarbamoyl, phenylcarbamoyl, benzylcarbamoyl,2-chloroethyl, 2-fluoroethyl, 2-bromoethyl, 2-hydroxyethyl,3-hydroxypropyl, 2-methoxyethyl, 2-ethoxyethyl, 2-methylthio-ethyl,2-methanesulphinylethyl, 2-methanesulphonyl-ethyl, 2-aminoethyl,3-aminopropyl, 2-methylamino ethyl, 2-dimethylaminoethyl, cyanomethyl,2-cyanoethyl, azidomethyl, 2-azidoethyl, ureidomethyl,3-amino-3-carboxypropyl, 2-(amidino)ethyl, 2-(N-aminoamidino)-ethyl,tetrahydropyran-2-yl, thietan-3-yl, 2-oxopyrrolidinyl and2-oxotetrahydrofuran-3-yl,

or, when R⁷ is of the formula XIV in which q is 1 or 2, a particularmeaning for R⁷ is when R⁸ and R⁹ are hydrogen or methyl,

or, when R⁷ is of the formula XV, a particular meaning for R⁷ is whenr=0 and R¹⁰ is hydrogen, methyl or methylthio, R¹¹ is hydrogen, methyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyano, carboxy,carboxymethyl, 2-carboxyethyl or methanesulphonylamino, or when R¹⁰ andR¹¹ are joined to form, together with the carbon to which they areattached, a cyclopropane, cyclobutane, cyclopentane, cyclohexane orcycloheptane ring and R¹² is hydroxy, amino, methoxy, ethoxy,methylamino, ethylamino, or of the formula NHOR¹³ in which R¹³ ishydrogen, methyl or ethyl.

Preferably R⁷ is C₁₋₆ alkyl for example methyl or ethyl,1-carboxycyclobutyl, 1-carboxycyclopentyl, or 2-carboxyprop-2-yl. Inparticular R⁷ is 2-carboxyprop-2-yl.

Particular meanings for R¹⁴ are hydrogen, methyl, ethyl or chlorine.

The cephalosporin derivatives referred to herein are generally named inaccordance with the `cephem` nomenclature and numbering system proposedin J.A.C.S. 1962, 84, 3400.

It will be realised, of course, that the present invention covers allisomeric and tautomeric forms of the aforementioned compounds. Forexample the rings of the formula (III) may be in pyranone orhydroxypyridine form.

As stated hereinbefore the compounds of this invention ar primarilyintended for use in therapy. Therefore in a preferred aspect the presentinvention provides a cephalosporin compound having a 3-positionsubstituent of the formula I or a pharmaceutically acceptable salt orester thereof. Suitable salts include acid addition salts such ashydrochloride, hydrobromide, citrate, maleate and salts formed withphosphoric and sulphuric acid. In another aspect suitable salts are basesalts such as an alkali metal salt for example sodium or potassium, analkaline earth metal salt for example calcium or magnesium, an organicamine salt for example triethylamine, morpholine, N-methylpiperidine,N-ethylpiperidine, procaine, dibenzylamine, or N,N-dibenzylethylamine.

In order to use a compound of the present invention or apharmaceutically acceptable salt or ester thereof for the therapeutictreatment of mammals including humans, in particular in treatinginfection, it is normally formulated in accordance with standardpharmaceutical practice as a pharmaceutical composition.

Therefore in another aspect the present invention provides apharmaceutical composition which comprises a cephalosporin compoundhaving a 3-position substituent of the formula I or a pharmaceuticallyacceptable salt or ester thereof and a pharmaceutically acceptablecarrier.

The pharmaceutical compositions of this invention may be administered instandard manner for the disease condition that it is desired to treat,for example by oral, rectal or parenteral administration. For thesepurposes it may be formulated by means known to the art into the formof, for example, tablets, capsules, aqueous or oily solutions orsuspensions, emulsions, dispersible powders, suppositories and sterileinjectable aqueous or oily solutions or suspensions.

In addition to the pharmaceutically acceptable cephalosporin derivativeof the present invention the pharmaceutical composition of the inventionmay also contain, or be co-administered with, one or more known drugsselected from other clinically useful antibacterial agents (for exampleother beta-lactams or aminoglycosides), inhibitors of beta-lactamase(for example clavulanic acid), renal tubular blocking agents (e.g.probenicid) and inhibitors of metabolising enzymes (for exampleinhibitors of peptidases, for example Z-2-acylamino-3-substitutedpropenoates).

A preferred pharmaceutical composition of the invention is one suitablefor intravenous, subcutaneous or intramuscular injection, for example asterile injectable containing between 1 and 50% w/w of the cephalosporinderivative, or one suitable for oral administration in unit dosage form,for example a tablet or capsule which contains between 100 mg. and 1 g.of the cephalosporin derivative.

The pharmaceutical compositions of the invention will normally beadministered to man in order to combat infections caused by bacteria, inthe same general manner as that employed for cephalothin, cefoxitin,cephradine, ceftazidime and other known clinically used cephalosporinderivatives, due allowance being made in terms of dose levels for thepotency of the cephalosporin derivative of the present inventionrelative to the known clinically used cephalosporins. Thus each patientwill receive a daily intravenous, subcutaneous or intramuscular dose of0.05 to 30 g., and preferably 0.1 to 10 g., of the cephalosporinderivative, the composition being administered 1 to 4 times per day,preferably 1 or 2 times a day. The intravenous, subcutaneous andintramuscular dose may be given by means of a bolus injection.Alternatively the intravenous dose may be given by continuous infusionover a period of time. Alternatively each patient will receive a dailyoral dose which is approximately equivalent to the daily parenteraldose. Thus a preferred daily oral dose is 0.5 to 10 g. of thecephalosporin derivative, the composition being administered 1 to 4times per day.

In a further aspect the present invention provides a process forpreparing a cephalosporin compound having a 3-position substituent ofthe formula I, which process comprises:

(a) reacting a cephalosporin compound having a 3-position substituent ofthe formula: --CH₂ L wherein L is a leaving group, with a source of--S--Q--(Y)_(n) --P wherein Y, Q, P and n are as hereinbefore defined;

(b) reacting a cephalosporin compound having a 3-position substituent ofthe formula, --CH₂ SH with a source of --Q--(Y)_(n) --P wherein Q, Y, Pand n are as hereinbefore defined;

(c) reacting a cephalosporin compound having a 3-position substituent ofthe formula --CH₂ --S--Q--J with a compound of the formula K--P whereinJ and K are such that the reaction takes place to form the link--(Y)_(n) -- between Q and P; or

(d) for preparing compounds of the formula (XIII), reacting a compoundof the formula (XVI) with a compound of the formula (XVII) or a reactivederivative thereof: ##STR7## wherein R⁴, R⁵, R⁶, X, Y, Q, P and n are ashereinbefore defined; or

e) for compounds of the formula (XIII) wherein R⁶ is a group ═NOR⁷,reacting a compound of the formula (XVIII): ##STR8## wherein R⁴, R⁵, X,Y, Q, P and n are as hereinbefore defined, with a compound of theformula: R⁷ ONH₂ wherein R⁷ is as hereinbefore defined; or

f) for compounds of the formula (XIII) wherein R⁶ is a group ═NOR⁷ andR⁷ is other than hydrogen, reacting a compound of the formula (XIII) ashereinbefore defined wherein R⁶ is a group ═NOH with a compound of theformula (XIX):

    L.sup.1 --R.sup.15                                         (XIX)

wherein L¹ is a leaving group and R¹⁵ is a group R⁷ other than hydrogen;or

g) for compounds of the formula (XIII) forming a group R⁵ by cyclisingan appropriate precursor thereof:

wherein any functional groups are optionally protected: and thereafter,if necessary:

i) removing any protecting group,

ii) for preparing in vivo hydrolysable esters, esterifying correspondinghydroxy groups,

iii) converting compounds wherein X is S to compounds wherein X issulphinyl and vice versa,

iv) forming a pharmaceutically acceptable salt.

In the reaction between a cephalosporin compound having a 3-positionsubstituent of the formula: --CH₂ L and a source of --S--Q--(Y)_(n) --P,conveniently L is a leaving group such as halo for example iodo, bromoor chloro, or is C₁₋₄ alkanoyloxy for example acetoxy. Typically asource of --S--Q--(Y)_(n) --P is the mercapto derivative HS--Q--(Y)_(n)--P. Such compounds are either known or are prepared in conventionalmanner by known methods as will be apparent to the skilled man. Inparticular reference should be made to the established techniques ofdisplacing leaving groups at the 3'-position of cephalosporinderivatives by a wide variety of sulphur nucleophiles. In a similarmanner a cephalosporin compound having a 3-position substituent of theformula --CH₂ SH is reacted with a source of --Q--(Y)_(n) --P, forexample L'--Q--(Y)_(n) --P wherein L' is a leaving group.

The cephalosporin starting materials for these reactions are known fromthe art, or are made by methods analogous thereto. See for exampleEP-A-127992 and EP-A-164944.

The reaction between compounds of the formulae (XVI) and (XVII) isperformed under conditions conventional in the cephalosporin art, forexample under standard acylation conditions wherein for example the acidis activated as an acid bromide, acid chloride, anhydride or activatedester, or the reaction is performed in the presence of a couplingreagent such as dicyclohexylcarbodi-imide.

The compounds of the formula (XVI) can be prepared in a manner analogousto that described for the compounds of the formula (I), with the 7-aminogroup being optionally protected.

The reaction between compounds of the formula (XVIII) and R⁷ ONH₂ isperformed under conditions standard in the general chemical and/orcephalosporin art. The compounds of the formula (XVIII) can be preparedin a manner analogous to that described for the compounds of the formula(I).

The reaction between the compound of the formula (XIII) wherein R⁶ is agroup ═NOH and a compound of the formula (XIX) is performed underconditions standard in the general chemical and/or cephalosporin art.

A group R⁵ may be formed by cyclizing an appropriate precursor. Forexample compounds of the formulae (XX) and (XXI): ##STR9## wherein R⁴,R⁶, X, Y, Q, P and n are as hereinbefore defined and L² is a leavinggroup, may be reacted to form a 2-aminothiazol-4-yl group. A nitrogenatom of the thiourea may be optionally protected during thiscyclization.

The compounds of the formula (XX) can be prepared in a manner analogousto that described for the compounds of the formula I.

The compounds of the formulae (XVII), (XIX) and R⁷ ONH₂ are known from,or can be made by the methods of, the general chemical and/orcephalosporin art.

The compounds of the formulae (XVI), (XVIII) and (XX) are novel and assuch form a further aspect of the present invention.

In the process of this invention any functional group can be optionallyprotected, if appropriate. Such protecting groups may in general bechosen from any of the groups described in the literature or known tothe skilled chemist as appropriate for the protection of the group inquestion, and may be introduced by conventional methods.

Protecting groups may be removed by any convenient method as describedin the literature or known to the skilled chemist as appropriate for theremoval of the protecting group in question, such methods being chosenso as to effect removal of the protecting group with minimum disturbanceof groups elsewhere in the molecule.

Specific examples of protecting groups are given below for the sake ofconvenience, in which "lower" signifies that the group to which it isapplied preferably has 1-4 carbon atoms. It will be understood thatthese examples are not exhaustive. Where specific examples of methodsfor the removal of protecting groups are given below these are similarlynot exhaustive. The use of protecting groups and methods of deprotectionnot specifically mentioned is of course within the scope of theinvention.

A carboxyl protecting group may be the residue of an ester-formingaliphatic or araliphatic alcohol or of an ester-forming phenol, silanolor stannanol (the said alcohol, phenol, silanol or stannanol preferablycontaining 1-20 carbon atoms).

Examples of carboxyl protecting groups include straight or branchedchain (1-12C)alkyl groups (eg isopropyl, t-butyl); halo lower alkylgroups (eg 2-iodoethyl, 2,2,2-trichloroethyl); lower alkoxy lower alkylgroups (eg methoxymethyl, ethoxymethyl, isobutoxymethyl); loweraliphatic acyloxy lower alkyl groups, (eg acetoxymethyl,propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl); loweralkoxycarbonyloxy lower alkyl groups (eg 1-methoxy-carbonyloxyethyl,1-ethoxycarbonyloxyethyl); aryl lower alkyl groups (eg p-methoxybenzyl,o-nitrobenzyl, p-nitrobenzyl, benzhydryl and phthalidyl); tri(loweralkyl)silyl groups (eg trimethylsilyl and t-butyldimethylsilyl);tri(lower alkyl)silyl lower alkyl groups (eg trimethylsilylethyl); and(2-6C)alkenyl groups (eg allyl and vinylethyl).

Methods particularly appropriate for the removal of carboxyl protectinggroups include for example acid-, base-, metal- or enzymically-catalysedhydrolysis.

Examples of hydroxyl .protecting groups include lower alkanoyl groups(eg acetyl); lower alkoxycarbonyl groups (eg t-butoxycarbonyl); halolower alkoxycarbonyl groups (eg 2-iodoethoxycarbonyl,2,2,2-trichloroethoxycarbonyl); aryl lower alkoxycarbonyl groups (egbenzoyloxycarbonyl, p-methoxybenzyloxycarbonyl,o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl); tri loweralkylsilyl (eg trimethylsilyl, t-butyldimethylsilyl) and aryl loweralkyl (eg benzyl) groups. In addition two hydroxy groups substituted onadjacent carbon atoms, for example in the catechol moiety, may beprotected in the form of a cyclic acetal such as the methylenedioxymoiety.

Examples of amino protecting groups include formyl, aralkyl groups (egbenzyl and substituted benzyl, eg p-methoxybenzyl, nitrobenzyl and2,4-dimethoxybenzyl, and triphenylmethyl); di-p-anisylmethyl andfurylmethyl groups; acyl (eg alkoxycarbonyl and aralkoxycarbonyl egt-butoxycarbonyl and benzyloxycarbonyl); trialkylsilyl (egtrimethylsilyl and t-butyldimethylsilyl); alkylidene (eg methylidene);benzylidene and substituted benzylidene groups, and the phthalimidogroup.

Esterification of hydroxy groups (ie R¹ and R²) to form in vivohydrolysable esters is performed in conventional manner. Reduction of acephalosporin sulphoxide to a cephalosporin and oxidation of asulphoxide to a sulphide are performed according to methods known in theart.

The following biological test methods, data and Examples serve toillustrate this invention.

ANTIBACTERIAL ACTIVITY

The pharmaceutically acceptable cephalosporin compounds of the presentinvention are useful antibacterial agents having a broad spectrum ofactivity in vitro against standard laboratory microorganisms, bothGram-negative and Gram-positive, which are used to screen for activityagainst pathogenic bacteria. The antibacterial spectrum and potency of aparticular compound may be determined in a standard test system. Thecompounds have particularly high activity in vitro against strains ofPseudomonas aeruginosa.

The antibacterial properties of the compounds of the invention may alsobe demonstrated in vivo in conventional mouse protection tests.

Cephalosporin derivatives have generally been found to be relativelynon-toxic to warm-blooded animals, and this generalisation holds truefor the compounds of the present invention. Compounds representative ofthe present invention were administered to mice at doses in excess ofthose required to afford protection against bacterial infections, and noovert toxic symptoms or side effects attributable to the administeredcompounds were noted.

The following results were obtained for representative compounds on astandard in vitro test system using Isosensitest agar medium. Theantibacterial activity is described in terms of the minimum inhibitoryconcentration (M1C) determined by the agar-dilution technique with aninoculum size of 10⁴ CFU/spot.

    ______________________________________                                                   MlC (μl/ml)                                                                EXAMPLE                                                            ORGANISM     1          5       9                                             ______________________________________                                        P. aeruginosa                                                                              0.06       0.25    0.125                                         PU21 (A8101028)                                                               Ent. cloacae 0.125      0.03    1                                             P99 (A8401054)                                                                Serr. marcesens                                                                            0.03       0.015   0.25                                          (A9421003)                                                                    Pr. morganii 0.03       0.125   0.5                                           (A8433001)                                                                    Kleb. aerogenes                                                                            0.015      0.008   0.125                                         (A8391027)                                                                    E. coli      0.008      0.008   0.03                                          DCO (A8341098)                                                                St. aureus   4          2       >128                                          147N (A8601052)                                                               S. dublin    0.03       0.008   0.125                                         (A8369001)                                                                    Strep. pyogenes                                                                            --         0.03    0.5                                           (A681018)                                                                     ______________________________________                                    

In the following Examples the following abbreviations are used:

7-ACA=7-aminocephalosporanic acid

AcOH=acetic acid

BSA=bistrimethylsilylacetamide

DMF=dimethylformamide

DMSO=dimethylsulphoxide

EtOH=ethanol

HPLC=high pressure liquid chromatography

MeOH=methanol

TEA=triethylamine

TFA=trifluoroacetic acid

The NMR spectra are taken at 90 MHz and are quoted in terms of deltavalues in parts per million (ppm) with reference to tetramethylsilane(delta=0). The solvent used was DMSOd₆ /CD₃ COOD/TFA except whereotherwise indicated. In the quotation of NMR data s=singlet, d=doublet,t=triplet, q=quartet, m=multiplet, br=broad.

EXAMPLE 17-[2-(2-Aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-[2-(3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid

(a) Methyl 3,4-dihydroxybenzoate (6 g) was heated with hydrazine hydrate(3.4 ml) at 110° C. with stirring for 15 minutes. The solid formed oncooling was triturated with ethanol and the crystals filtered to yield 5g (83%) of the hydrazide of 3,4-dihydroxybenzoic acid, NMR (DMSO_(d6)+AcOD): 6.7 (d,1H); 7.2 (dd,1H); 7.3 (d,1H).

(b) To the product from (a) above (4.5 g) in absolute ethanol was addedKOH (5.6 g) and carbon disulphide (2.4 ml). Stirring was continued for20 hours. The solvents were evaporated, and the crude product (9 gpotassium 3,4-dihydroxy-benzoyldithiocarbazate) used in the next step.

(c) The product of (b) above was added in small portions, with stirring,to conc. H₂ SO₄ (40 ml) cooled below 5° C., over 30 minutes. The crudeproduct was poured on to crushed ice and the precipitate filtered andwashed with water. It was then dissolved in ether and washed toneutrality with water. The ether was evaporated and the solid dissolvedin dichloromethane, insoluble material was filtered off afterevaporation of dichloromethane and2-mercapto-5-(3,4-dihydroxyphenyl)-1,3,4-thiadiazole (1.2 g) obtained,NMR (DMSO_(d6), AcOD, TFA): 6.84 (d,1H); 7.04 (dd,1H); 7.15 (d,1H).

(d)7-[2-(2-Aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-acetoxymethylceph-3-em-4-carboxylicacid (527 mg), the product of (c) above (270 mg) and sodium iodide (1.5g) in 5 ml water at pH 6.5-7 were heated at 65° C. for 5.5 hours. The pHwas maintained at 7 by the addition of AcOH or NaHCO₃ during thereaction, which was followed by HPLC. When reaction was complete, thesolvent was evaporated and the crude product purified by preparativeHPLC eluting with MeOH/(NH₄)₂ CO₃ buffer 30/70 to yield the titlecompound (58 mg), NMR (DMSO_(d6), AcOD, TFA): 1.56 (s,6H); 3.7 (m,2H);4.4 (m,2H); 5.2 (d,1H); 5.9 (d,1H); 6.85 (d,1H); 7.05 (s,1H); 7.1(dd,1H); 7.35 (d,1H).

EXAMPLE 2 7-[2-(2-Aminothiazol-4-yl)-2-((Z)-ethoxyimino)acetamido]-3-[2-(3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid

1 equivalent of7-amino-3-[2-(3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid was silylated with 4 equivalents of bistrimethylsilylacetamide inanhydrous dichloromethane at room temperature for three hours. To thissolution, cooled to 0° C. under argon, was added 1 equivalent of2-[(Z)-ethoxyimino-2-(tritylaminothiazol-4-yl) acetic acid chloride(prepared from the corresponding acid (1 equivalent), 1 equivalent PCl₅and 1 equivalent TEA in dichloromethane at 0° C. for 2 hours followed byevaporation of the solvent and all POCl₃, and redissolution indichloromethane). After a few minutes at 0° C., the solvent wasevaporated, and all the protecting groups removed by treating the crudereaction mixture with TFA/water for 1 hour at room temperature. Thesolvents were evaporated and the crude product dissolved in MeOH andprecipitated in ether. Further purification was achieved by MPLC, toyield the title compound (7%), NMR (DMSO_(d6), AcOD, TFA), 1.25 (t,3H);3.7 (m,2H); 4.2 and 4.25 (2d,2H); 4.4 (q,2H); 5.15 (d,1H); 5.75 (d,1H);6.85 (d,1H); 7 (s,1H); 7.15 (d,1H); 7.3 (s,1H).

EXAMPLE 37-[1-(2-Aminothiazol-4-yl)-1-(Z)-propenecarboxamido]-3-[(2-(3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid

To a solution of (Z)-1-(2-aminothiazol-4-yl)-propene-1-carboxylic acid(370 mg, 2 mmol) (prepared as described in European Patent ApplicationNo. 0107138) and N-ethyl-diisopropylamine (380 μl, 2.2 mmol) in DMF (3ml) at -50° C. was added methylsulphonyl chloride (CH₃ SO₂ Cl) (170 μl,2 mmol). The mixture was stirred for 1 hour at -50° C. The solution wasthen added rapidly to a cold (0° C.) solution of7-amino-3-[(2-(3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid (300 mg, 2 mmol) and TEA (560 μl, 4 mmol in 2 ml DMF and 0.5 mlwater). Stirring was continued at room temperature for 30 minutes. Thesolvents were evaporated and the product purified by preparative HPLC,eluting with MeOH/(NH₄)₂ CO₃ buffer 30/70 to yield the title compound(50 mg), NMR (300 MHz) (DMSO_(d6) , AcOD, TFA): 1.8 (d,3H), 3.6 (m,2H),4-4.5 (m,2H); 5.1 (d,1H); 5.7 (d,1HO); 6-6.4 (m,1H), 6.8 (s,1H), 6.7-7.3(m,3H).

PREPARATION OF STARTING MATERIAL7-Amino-3-[(2-(3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid.

(Starting material for Examples 2 and 3) was prepared as follows:

To a solution of 2-mercapto-5-(3,4-dihydroxyphenyl)-1,3,4-thiadiazole(695 mg) in water (10 ml) and DMF (3 ml) was added NaHCO₃ (1.03 g)followed by 7-amino-3-iodomethylceph-3-em-4-carboxylic acid (1.2 g).Stirring was continued for 2 hours at room temperature. The mixture wasacidified to pH 3.5-4 and the title compound (1.1 g) was obtained as aprecipitate, which was filtered off and dried. NMR (DMSO_(d6), AcOD,TFAd) 3.75 (m,2H) 4.2-4.6 (m,2H), 5.1 (d,1H); 5.2 (d,1H), 6.85 (d,1H);7.15 (d,1H); 7.3 (d,1H).

EXAMPLE 4

The process of Example 3 was repeated starting from(Z)-1-(2-aminothiazol-4-yl)butene-1-carboxylic acid (prepared asdescribed in European Patent Application No. 0107138 which comprisesabout 25% (E)-isomer and about 75% (Z)-isomer) to form the (E)- and(Z)-isomers of7-(1-(2-aminothiazol-4-yl)-1-(Z)-butenecarboxamido]-3-[(2-3,4-dihydroxyphenyl)-1,3,4-thiadiazol-5-ylthiomethyl]ceph-3-em-4-carboxylicacid. The isomers of the final product were separated using preparativeHPLC using ammonium phosphate buffer/MeOH of 40% MeOH increasing to 50%yielding (Z)-isomer (190 mg; 16%) and (E)-isomer (27 mg; 3%) having NMRas follows in DMSO_(d6), AcOD, TFA:

(Z)-isomer (300 MHz):

1.05 (l,3H); 2.25 (m,2H); 3.65 (d,1H); 3.8 (d,1H); 4.25 (d,1H); 4.55(d,1H); 5.15 (d,1H); 5.75 (d,1H); 6.25 (t,1H); 6.55 (s,1H); 6.85 (d,1H);7.15 (dd,1H); 7.35 (d,1H).

(E)-isomer (90 MHz):

1.05 (t,3H); 2.2 (m,2H); 3.7 (m,2H); 4.24 (d,1H); 4.52 (d,1H); 5.1(d,1H); 5.7 (d,1H); 6.7-7.4 (m,5H).

EXAMPLE 57-[2-(2-Aminothiazol-4-yl)-2-((Z)-methoxyimino)acetamido]-3-[(6,7-dihydroxyquinazolin-4-yl)thiomethyl]ceph-3-em-4-carboxylicacid. ##STR10##

(a) (1) (5 g) (prepared from the corresponding oxo compound usingLawesson's reagent in acetonitrile) was dissolved in dichloromethane(anhydrous) (75 ml) with the addition of BSA (17 ml). BBr₃ (13 ml) wasadded to the solution at 0° C. The reaction mixture was stirred at roomtemperature overnight. The dichloromethane was evaporated and theresidue dissolved in MeOH, evaporated, and purified by HP20SSchromatography. The dihydroxy product (2.6 g) was obtained: NMR(DMSO_(d6) /TFA); 7.13 (s,1H); 7.92 (s,1H); 8.91 (s,1H).

(b) 7-Aminocephalosporanic acid (1.4 g) in acetonitrile (10 ml) andboron trifluoride ethoxylate (4 ml) was treated with (2) (1 g) at roomtemperature for 2.5 hours. Acetonitrile was evaporated, the crudereaction mixture poured into water, and the precipitate filtered anddried. A crude product (1.9 g) was obtained, which was purified bychromatography over HP20SS resin, eluting with water containing 1% AcOH,and MeOH at varying proportions. Two cephalosporin derivatives were thusobtained. The more polar derivative (3) (280 mg) eluted at 5% MeOH andthe less polar derivative (330 mg) eluted at 10% MeOH (330 mg). NMR(DMSO_(d6), TFA) (more polar derivative): 3.64 (d,1H); 3.88 (d,1H); 4.32(d,1H); 4.96 (d,1H); 5.2 (s,2H); 7.37 (s,1H); 7.45 (s,1H); 9.11 (s,1H).

(c) The cephalosporins (3) (200 mg), mercaptobenzthiazolyl2-(2-aminothiazol-4-yl)-2-methoxyiminoacetate (174 mg) and Et₃ N (35 μl)in DMF (3 ml) were stirred at room temperature for 2 hours. The solventwas then evaporated and the product purified by chromatography overHP20SS resin, eluting with ammonium carbonate buffer/MeOH usingincreasing amounts of MeOH. The title compound (39 mg) was obtained, NMR(DMSO_(d6), TFA, AcOH): 3.55 (d,1H) 3.85 (d,1H); 3.95 (s,3H); 4.25(d,1H); 4.95 (d,1H); 5.15 (d,1H); 5.75 (d,1H); 6.95 (s,1H); 7.35 (s,1H);7.45 (s,1H); 9.05 (s,1H).

EXAMPLE 67-[2-(2-Aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-[6,7-dihydroxyquinazolin-4-ylthiomethyl]ceph-3-em-4-carboxylicacid. ##STR11##

(a) The thioester (A) (400 mg) was dissolved in dichloromethane (2 ml)at room temperature and trimethylsilyl iodide (250 μl) added. Themixture was stirred at room temperature for 4 hours under argon toobtain a solution of the silylated compound (B) in dichloromethane.

(b) Cephalosporin (C) (106 mg) (obtained as in Example 5(b)) indichloromethane (2 ml) was stirred for one hour at 40° C. under argonwith BSA (514 μl) thus forming a solution of the silylated cephalosporin(D) in dichloromethane.

(c) The solution of (D) obtained in (b) was added to the solution of (B)obtained in (a) and stirring was continued for 2 hours at roomtemperature. The solvents were then evaporated, and the crude productpurified by HPLC using ammonium carbonate/MeOH as eluant, withproportions of MeOH varying from 20 to 35%, to yield the title compound(48 mg), NMR (DMSO_(d6), AcOD, TFA): 1.5 (s,6H); 3.52 (d,1H); 3.8(d,1H); 4.24 (d,1H); 4.92 (d,1H); 5.16 (d,1H); 5.84 (d,1H); 7.0 (s,1H);7.32 (s,1H); 7.4 (s,1H); 9.04 (s,1H).

EXAMPLE 77-[2-(2-Aminothiazol-4-yl)-2-((Z)-methoxyimino)acetamido]-3-[(1-(2-(3,4-dihydroxyphenyl)ethyl)tetrazol-5-yl)thiomethyl]-ceph-3-em-4-carboxylicacid. ##STR12##

(a) Dopamine hydrochloride (3.8 g) was dissolved in EtOH (40 ml) and Et₃N (4.04 g) added at room temperature followed by CS₂ (1.52 g). Themixture was stirred at room temperature for 2 hrs. until all of thestarting material had disappeared (by tlc); to form (1).

(b) Methyl iodide (2.84 g) was added to the product of (a) in solution,stirring for 2 hours at room temperature. The solvents were evaporated,and the crude product purified by silica gel chromtography eluting withdichloromethane/ether to yield the thioester (2) (4.3 g), NMR(DMSO_(d6)) 2.5 (s,3H); 2.7 (t,2H); 3.5-3.9 (m,2H); 6.3-6.7 (m,3H); 8.56(s,1H); 8.68 (s,2H); 9.9 (t,1H).

(c) The thioester (2) (4 g) was dissolved in water (20 ml) and EtOH (20ml). NaN₃ (1.6 g) was added to the solution, which was then refluxed for1 hour 45 mins. The solvents were evaporated, and the crude productpurified by silica gel chromatography eluting with CH₂ Cl₂ --MeOH(100→50--50) to obtain compound (3) (1.49 g), NMR (DMSO_(d6)): 2.9(t,2H); 4.3 (t,2H); 6.3-6.7 (m,3H).

(d) Compound (3) (238 mg) was condensed with 7-ACA (272 mg) in CH₃ CN (3ml) with BF₃ /Et₂ O (1.3 ml) at 40° for 30 mins. The solvents wereevaporated and the mixture purified by HP₂₀ SS chromtography elutingwith MeOH/H₂ O using a gradient of MeOH. The cephalosporin (4) wasobtained, NMR (DMSO_(d6), ACOD, TFA) 2.94 (t,2H); 3.7 (s,2H); 4.1-4.5(m,4H); 5.1 (s,2H); 6.2-6.7 (m,3H).

(e) The cephalosporin (4) (120 mg) was dissolved in DMF (3 ml) and Et₃ N(26 mg) added, followed by thioester (5) (91 mg). The mixture wasstirred at room temperature for 2.5 hours. The solvents were evaporatedand the crude product purified using HP20SS chromtography eluting withH₂ O/MeOH with a gradient of MeOH to give the title compound (120 mg),NMR (DMSO_(d6), AcOD, TFA): 2.92 (t,2H); 3.52 (d,1H); 3.76 (d,1H); 3.96(s,3H); 4.1-4.6 (m,4H); 5.1 (d,1H); 5.76 (d,1H); 6.2-7 (m,4H).

EXAMPLE 8 7-[2-(2-Aminothiazol-4-yl)1-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-[1-(2-carboxy-2-(3,4-dihydroxyphenyl)ethyl)tetrazol-5-ylthiomethyl]-ceph-3-em-4-carboxylicacid. ##STR13##

a) To a suspension of D,L-Dopa (5 g) in ethanol (50 ml) was addedtriethylamine (7 ml), followed by CS₂ (1.5 ml). The mixture was stirredat room temperature for 2 hours, water (20 ml) was added and stirringwas continued for a further 12 hours. Methyliodide (1.55 ml) was added,the mixture was stirred for a further 2 hours, the solvents wereevaporated and the mixture was subjected to chromatography on silicaeluting with CH₂ Cl₂ /CH₃ OH (100:0→70:30) to give the thioester (3)(6.26 g), NMR (DMSO-d₆ /TFAd) 2.53 (s,3H); 3.14 (d,2H); 5.1 (t,1H);6.4-6.8 (m,3H).

b) To the thioester (3) (3.54 g) in methanol (80 ml) and acetonitrile(80 ml) was added p-toluenesulphonic acid (1.25 g) and then, slowly, wasadded an excess of diazodiphenylmethane. This mixture was stirred at 40°C. for 3 hours and at room temperature for a further 16 hours. Thesolvents were evaporated, the mixture dissolved in dichloromethane andpurified by chromatography over silica eluting with CH₂ Cl₂ /CH₃ OH(100:0→90:10) to give the ester (4) (2.8 g), NMR (CDCl₃) 2.59 (s,3H);3.10-3.3 (m,2H); 4.7-5.2 (m,1H); 6.13 (d,1H); 6.29 (dd,1H); 6.56 (d,1H);6.93 (s,1H); 7.31 (2s,1OH).

c) To the ester (4) (2.66 g) in ethanol (10 ml) was added sodium azide(565 mg) in water (5 ml) at room temperature. The mixture was stirredunder reflux for 1 hour, the solvents were evaporated and the residuepurified by chromatography over silica eluting with CH₂ Cl₂ /CH₃ OH(100:0→60:40) to give compound (5) (1.5 g), NMR (DMSO-d₆) 3.41 (d,2H);5.80 (t,1H); 6.46, 6.54, 6.61 (3s,3H); 6.8 (s,1H); 7.31 (s,10H).

d) The compound (5) (830 mg) was dissolved in trifluoroaceticacid/anisole at room temperature. After 40 minutes the solvents wereevaporated and the crude acid (6) was obtained. This acid (6) (518 mg)and 7-ACA (489 mg) were suspended in acetonitrile (6 ml) and BF₃ Et₂ O(2.4 ml) was added with stirring. The mixture was stirred for 35 minutesat 40° C., the solvents were evaporated, and the residue was trituratedunder ether to give a solid which was purified on HP20SS resin elutingwith H₂ O/CH₃ OH (100:0→90:10) to give the cephalosporin (7) (497 mg),NMR (DMSO-d₆ /CD₃ COOD) 3.3 (d,2H); 3.67 (s,2H); 4.16, 4.48 (2d,2H);5.13 (s,2H); 5.48 (t,1H); 6.2-6.6 (m,3H).

e) To the cephalosporin (7) (165 mg) in DMF (4 ml) was addedtriethylamine (46 μl) and compound A (159 mg) (as pictured in Example6). The mixture was stirred at room temperature for 2.5 hours, thesolvents were evaporated and the residue was treated with TFA (4 ml) atroom temperature for 1 hour. TFA was evaporated and the residue purifiedby HPLC eluting with H₂ O/AcOH/CH₃ OH (80:1:20→65:1:35) to give thetitle compound (111 mg), NMR (DMSO-d₆ /CD₃ COOD/TFA-d) 1.53 (s,6H); 3.38(d,2H); 3.6 (s,2H); 4.16-4.18 (2d,2H); 5.12 (d,1H); 5.84 (d,1H); 5.5(t,1H); 6.2-6.7 (m,3H); 7.04 (s,1H).

EXAMPLE 97-[2-(2-Aminothiazol-4-yl)-2-((Z)-carboxy-1-methylethoxyimino)acetamido]-3-[(1-(2-carboxymethyl-4,5-dihydroxyphenyl)tetrazol-5-yl)thiomethyl]-ceph-3-em-4-carboxylic acid. ##STR14##

a) 3,4-Dimethoxyphenylacetic acid (7.84 g) was solubilised in ether (100ml) and AcOH (20 ml). Fuming HNO₃ (1.68 ml) was added to the solutiondropwise. Stirring was maintained at room temperature for 16 hours. Theprecipitate was filtered, washed with ether and dried to give compound(1) (4.19 g), NMR (DMSO-d₆) 3.88 (s,3H); 3.80 (s,3H); 3.97 (s,2H); 7.15(s,1H); 7.70 (s,1H).

b) Compound (1) (4.19 g) was solubilised in methanol (200 ml) and TFA (2ml). 10% Pd/C (200 mg) was added and the mixture was hydrogenated atroom temperature and atmospheric pressure for 2 hours. The catalyst wasfiltered off and the solvents evaporated to give (2) as a crude residue.

c) Compound (2) (3.6 g) was solubilised in actonitrile (35 ml) and water(35 ml) in the presence of triethylamine (3.6 ml). The mixture wascooled to 0° C. in an ice-bath, thiophosgene (1.6 ml) was added dropwiseand after a few minutes the mixture was acidified with 2N HCL andextracted into ether to give isothiocyanate (3) (2.3 g). This wasdissolved in ethanol (60 ml) and water (30 ml) containing sodiumbicarbonate (830 mg). An aqueous solution of sodium azide (897 mg) inwater (30 ml) was added, the mixture was maintained at 40° C. for 31/2hours, cooled, acidified with 2N HCl, evaporated and the residue waspurified by HP20SS chromatography, eluting with H₂ O/AcOH(100:1:0→25:1:75) to give compound (4) (960 mg), NMR (DMSO-d₆) 3.51(s,3H); 3.78, 3.86 (2s,6H); 7.12 (s,2H).

d) To compound (4) (900 mg) in dichloromethane (50 ml) was added BSA(1.5 ml). The solution was cooled to 0° C. and BBr₃ (2 ml) added withstirring for 31/2 hours. The solvents were evaporated and the residuehydrolysed by pouring into water at 0° C. The resultant reaction mixturewas purified by chromatography on HP20SS resin eluting with H₂O/AcOH/CH₃ OH (100:1:0→80:1:20) to give the dihydroxy compound (5) (700mg), NMR 3.33 (s,2H); 6.79, 6.87 (2s,2H).

e) To dihydroxy compound (5) (650 mg) and 7-ACA (655 mg) in acetonitrile(25 ml) was added BF₃.Et₂ O (4 ml) at room temperature with stirring.After stirring at 40° C. for 30 minutes the solvent was evaporated andthe residue triturated under ether and purified by HP20SS resinchromatography eluting with H₂ O/AcOH/CH₃ OH (100:1:0→ 80:1:20) to givethe cephalosporin (6) (622 mg), NMR (DMSO-d₆ /TFAd) 3.22 (s,2H); 3.80(s,2H); 4.28 (d,1H); 5.19 (s,2H); 6.81, 6.92 (2s,2H).

f) To cephalosporin (6) (160 mg) in DMF (4 ml) was added triethylamine(46 μl) and Compound A (159 mg) (as pictured in Example 6). The mixturewas stirred for 4 hours at room temperature, the solvents wereevaporated and the residue dissolved in TFA (4 ml) and stirred for 21/2hours. The TFA was evaporated and the residue purified by HP20SS resinchromatorgraphy eluting with H₂ O/AcOH/CH₃ OH (100:1:0→40:1:60) to givethe title compound (120 mg), NMR (DMSO-d_(6/) TFAd) 1.53 (s,6H); 3.21(s,2H); 3.70 (s,2H); 4.20 (d,1H); 4.60 (d,1H); 5.16 (d,1H); 5.83 (d,1H);6.78, 6.90 (2s,2H); 7.05 (s,1H).

EXAMPLE 107-[2-(2-Aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido-3-[(1-(2-(3,4-dihydroxyphenyl)ethyl)tetrazol-5-yl)thiomethyl]ceph-3-em-4-carboxylic acid. ##STR15##

To cephalosporin (1) (135 mg) (see Example 7) in solution in DMF, wasadded Et₃ N (42 μl) followed by Compound A (174 mg) (as depicted inExample 6), at room temperature. Stirring was continued for 21/2 hours,the solvent was evaporated, the residue was dissolved in TFA (3 ml) andstirred at room temperature for 1 hour. The solvent was evaporated andthe residue chromatographed over HP20SS resin eluting with H₂ O/AcOH/CH₂OH (45.1:55) to give the title compound (98 mg) NMR (DMSOd₆, CD₃ COD,TFAd) 1.55 (s,6H); 2.95 (t,2H); 3.5-3.8 (m,2H); 4.1-4.6(m,4H); 5.15,6.85 (2d,2H); 6.2-6.7 (m,3H); 7.05 (s,1H).

We claim:
 1. A compound of the formula (XVI): ##STR16## wherein: X issulphur or sulphinyl;R⁴ is hydrogen, methoxy or formamido; wherein--S--Q-- is of the formula (VIII): ##STR17## which optionally may bear apositive charge, and which optionally may be substituted on an availablenitrogen atom by carboxy, sulpho, C₁₋₄ alkoxycarbonyl or C₁₋₄ alkyl(which alkyl group may itself optionally be substituted by carboxy,sulpho or C₁₋₄ alkoxycarbonyl); P represents: (i) a benzene ring(optionally fused to a further benzene ring (so forming a naphthylgroup) or to a 5 or 6 membered heterocyclic aromatic group containing 1,2 or 3 heteroatoms selected from nitrogen, oxygen and sulphur) saidbenzene ring (or in the case of naphthyl either benzene ring)substituted by groups R¹ and R² which are ortho with respect to oneanother wherein R¹ is hydroxy or an in vivo hydrolysable ester thereofand R² is hydroxy, an in vivo hydrolysable ester thereof, carboxy,sulpho, hydroxymethyl, methanesulphonamido or ureido; (ii) a group ofthe formula (II): ##STR18## or, (iii) a group of the formula (III):##STR19## wherein M is oxygen or a group NR³ ; wherein R³ is hydrogen orC₁₋₄ alkyl: ring P (or, in the case wherein ring P is a benzene ring andis fused to another benzene ring, either benzene ring) is optionallyfurther substituted by C₁₋₄ alkyl, halo, hydroxy, hydroxy C₁₋₄ alkyl,cyano trifluoromethyl, nitro, amino, C₁₋₄ alkylamino, di-C₁₋₄alkylamino, amino C₁₋₄ alkyl, C₁₋₄ alkylamino C₁₋₄ alkyl, di-C₁₋₄alkylamino C₁₋₄ alkyl, C₁₋₄ alkanoyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio, C₁₋₄alkanoyloxy, carbamoyl, C₁₋₄ alkylcarbamoyl, di-C₁₋₄ alkyl carbamoyl,carboxy, carboxy C₁₋₄ alkyl, sulpho, sulpho C₁₋₄ alkyl, C₁₋₄alkanesulphonamido, C₁₋₄ alkoxycarbonyl, C₁₋₄ alkanoylamino, nitroso,thioureido, amidino, ammonium, mono-, di- or tri-C₁₋₄ alkylammoniumpyridinium, or a 5-membered heterocyclic ring containing 1 to 4heteroatoms selected from oxygen, nitrogen and sulphur which isoptionally substituted by 1, 2 or 3 C₁₋₄ alkyl or C₁₋₄ alkoxy groups,n=0 or 1 such that when n=1 Y represents a covalent bond between Q and Por a (1-4C)alkylene group optionally substituted by carboxy or sulpho orY represents a group --(CH₂)_(m) --Y'-- wherein m=1 or 2 and Y' is--O.CO-- or --NH.CO--; and when n=0 Q and P both represent monocyclicrings which are fused on an available carbon--carbon or carbon--nitrogenbond.
 2. A compound according to claim 1 wherein P is a benzene ringsubstituted by groups R¹ and R² which are ortho to one another whereinR¹ and R² are independently hydroxy or an in vivo hydrolysable esterthereof said benzene ring being optionally further substituted.
 3. Acompound according to claim 1 wherein --(Y)_(n) -- represents a covalentbond between groups Q and P.
 4. A compound according to claim 1 wherein--(Y)_(n) -- is methylene or ethylene.